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Molecular cloning, polymorphism, and expression analysis of the LKB1/STK11 gene and its association with non-specific digestive disorder in rabbits

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Abstract

Liver kinase B1 (LKB1, also called STK11) encodes a serine/threonine kinase mutated in Peutz–Jeghers cancer syndrome characterized by gastrointestinal polyposis. Although LKB1 plays an important role in regulating energy homeostasis, cell growth, and metabolism via activation of adenosine monophosphate (AMP)-activated protein kinase (AMPK), nothing is known about its molecular characteristics and possible involvement in non-specific digestive disorder (NSDD) of rabbits. In the present study, we first cloned the coding sequence (CDS) of rabbit LKB1, which consisted of 1317 bp encoding 438 amino acids (AAs) and contained a highly conserved S_TKc kinase domain. Its deduced AA sequence showed 87.93–91.10% similarities with that of other species. In order to determine its involvement in NSDD, a NSDD rabbit model was built by a dietary fiber deficiency. The polymorphic site of LKB1 was then investigated in both healthy and NSDD groups using directing sequencing. Our results suggested that a synonymous variant site (840 c. G > C, CCC→CCG) existed in its S_TKc domain, which was associated with susceptibility to NSDD. Furthermore, qPCR was utilized to examine the mRNA levels of LKB1 and its downstream targets (i.e., PRKAA2, mTOR and NF-kβ) in several intestinal-related tissues from both healthy and NSDD groups. Significant changes in their expression levels between two groups indicated that impaired LKB1 signaling contributed to the intestinal abnormality in NSDD rabbits. Taken together, it could be concluded that LKB1 might be a potential candidate gene affecting the occurrence of rabbit NSDD. This information may serve as a basis for further investigations on rabbit digestive diseases.

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Acknowledgements

This study was financially supported by the 12th Five-Year Breeding Research projects in Sichuan (Grant No. 2011NZ0099-4), China Agricultural Research System (Grant No. CARS-44-A-2), and the double-support project of Sichuan Agricultural University.

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    1. Institute of Animal Genetics and Breeding, Sichuan Agricultural University, Chengdu Campus, Huimin Road #211, Wenjiang, 611130, Sichuan, China

      Yanhong Li, Shenqiang Hu, Jie Wang, Shiyi Chen, Xianbo Jia & Songjia Lai

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    Yanhong Li and Shenqiang Hu contributed equally to this work.

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    Li, Y., Hu, S., Wang, J. et al. Molecular cloning, polymorphism, and expression analysis of the LKB1/STK11 gene and its association with non-specific digestive disorder in rabbits. Mol Cell Biochem 449, 127–136 (2018). https://doi.org/10.1007/s11010-018-3349-1

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